How to Solve Uneven Distribution of Cells in A Cell Culture Plate
Many people use the cell culture plate for cell biology experiments, but there are often problems, such as cells are sometimes not very evenly distributed: either dense in the middle and sparse around or dense around and bald in the middle. Here are some experiences to share with you.
Causes and treatment measures of uneven cell distribution in cell culture plate:
- First of all, it is necessary to find out how the cells are mixed, whether it is blowing with a pipette or shaking the growth area cell culture plates.
- If it is the latter and shaken in a circle, it is very likely that thrown the cells to the surrounding part due to the centrifugal force, resulting in fewer cells in the middle.
- Of course, it is also possible that the quality of the growth area cell culture plates is a problem or the cell density is too low when plating.
- For this, before culturing the seed plate, put the culture plate into the incubator for a few hours of saturation and then take it out, when seeding the cells, with light force. You can use a dripper to slowly add it to the side of the culture plate hole.
- The cell suspension flows into the wells of the plate, and the cultured cells grow substantially uniformly. Remember to never shake with a shaker or your cells will clump together.
- There are also some feasible treatment methods: blow and beat evenly before adding; add slowly from the side or bottom of the perforated plate.
- Some people may think that the uneven coating of the petri dish will cause this phenomenon because when the mixed blood vessel segments are in the petri dish, the blood vessel segments are evenly in the petri dish under the microscope.
- In the middle, the vascular segments that adhered to the wall after 24 hours were more in the periphery, and relatively few in the middle.
Some tips about the polystyrene cell culture plate:
The difference between a cell culture plate and an ELISA plate:
- It is definitely possible to measure absorbance with a multi-well culture plate, and often for protein quantification and MTT detection of samples.
- Difference: ELISA plates are generally more expensive than polystyrene cell culture plates.
- Cell plates are mainly for cell culture, but they can also measure protein concentration; ELISA plates include coating plates and reaction plates, which are generally not for cell culture.
- Protein detection after immunoenzyme-linked reaction requires higher requirements and a specific enzyme-labeled working solution.
How to properly culture cells in a cell cultures plate:
- Attempt to digest and disperse cells into a single-cell suspension. For easy-to-digest cells, wash once with DPBS, add 0.5ml of trypsin to rinse once (25cm2 culture flask or 6-well plate), discard the trypsin, and digest at 37 degrees for 2-3 minutes or at room temperature for about 5 minutes.
- Observe under the microscope whether the cell digestion is more dispersed. If that’s not enough, extend the digestion time.
- Inoculated 96-well cell culture plates with 100 microliters per well. Aspirate the cell suspension directly with a 100 μl pipette and insert it directly along the wall of the well (a little further from the bottom of the well, do not make the bottom too high).
- Not pumped the dispenser to the maximum, press gently, replace the tip after each row, and gently mix the cell suspension.
- Close the lid when all is added, gently hold the left side of the plate with your left hand and tap the right edge of the plate with your right hand. Pay attention to grip strength.
- Too strong or too many times can cause the cells to concentrate. Stack, rotate the plate clockwise (counterclockwise is not good), tap the remaining three sides, in turn, let it stand for about 5 minutes, and put it into a 37-degree incubator.